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For the quantitative determination of alkaline phosphatase in human serum. Distributed in almost every tissue of the body, serum alkaline phosphatase (ALP) levels are of interest in the diagnosis of hepatobiliary disorder and bone disease. Most of the ALP in the normal adult serum is from the liver or biliary tract. Normal alkaline phosphatase levels are age dependent, and levels are elevated during periods of active bone growth. Moderate elevations of ALP (not involving the liver or bone) may be attributed to Hodgkin's disease, congestive heart failure, and abdominal bacterial infections. Elevations also occur in the third trimester of pregnancy. Alkaline phosphatase is determined by measuring the rate of hydrolysis of various phosphate esters. p-Nitrophenyl Phosphate is one such ester that was used as a substrate by Fujita in 1939. Bowers and McComb further modified the procedure to a kinetic assay. In 1974, the Committee on enzymes of the Scandinavian Society for Clinical Chemistry and Clinical Physiology adopted a modification of the above procedure as the recommended procedure. The present method is a modification of the reference methods of the above committee and the American Association for Clinical Chemistry.